Autoradiographic evidence of differential loss of BUdR-substituted DNA after uv exposure in FPG harlequin staining
| Autor: | C.H. Ockey |
|---|---|
| Rok vydání: | 1980 |
| Předmět: |
Hot Temperature
Staining and Labeling Differential staining Ultraviolet Rays Chinese hamster ovary cell Cell Biology DNA Biology Chromatids Staining technique Molecular biology Staining Cell Line chemistry.chemical_compound chemistry Biochemistry Bromodeoxyuridine Cricetinae Animals Autoradiography Chromatid Metaphase Uv treatment Thymidine |
| Zdroj: | Experimental cell research. 125(2) |
| ISSN: | 0014-4827 |
| Popis: | Autoradiographic analysis of CHO cells labelled with [3H]TdR or [3H]BUdR shows that only [3H]BUdR label is removed from metaphase chromosomes after FPG staining. [3H]BUdR is differentially removed from the bifilary labelled chromatid (BB) compared with the unifilary labelled chromatid (TB). UV treatment alone removes the label and produces harlequin staining and if the UV step is omitted from the FPG staining technique no loss of label or harlequin staining occurs. The heat treatment step (60 °C in 2 × SSC) removes further label, reducing the ratio of grains BB TB to 0.8:1.0 and improving the differential staining. Over-treatment with heat produces paler staining chromatids without altering this ratio. The differential loss of BUdR-substituted DNA through UV photolysis and extraction in solution appears to be the cause of the differential harlequin staining of chromatids in this technique. |
| Databáze: | OpenAIRE |
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